Home » Expanding the scope of MS binding assays to low-affinity markers as exemplified for mGAT1

TITLE

Expanding the scope of MS binding assays to low-affinity markers as exemplified for mGAT1

AUTHOR(S)

Zepperitz, Christine; Höfner, Georg; Wanner, Klaus T.

PUB. DATE

May 2008

SOURCE

Analytical & Bioanalytical Chemistry;May2008, Vol. 391 Issue 1, p309

SOURCE TYPE

Academic Journal

DOC. TYPE

Article

ABSTRACT

Following a recently developed concept of MS binding assays based on the quantification of a native marker by LCâ€“MS a procedure to study binding of a low-affinity marker in kinetic, saturation, and competition experiments was established. Separation of bound and unbound markerâ€”the most crucial step of the assayâ€”could be effectively achieved by filtration in a 96-well-format. MS binding assays according to this procedure allowed the reliable characterization of NO 711 binding to mGAT1 in presence of physiological NaCl concentrations. Comparing the results obtained in the present study with those from experiments using 1 mol Lâˆ’1 NaCl in the incubation milieu reveals remarkable differences with respect to the markerâ€™s affinity and kinetics and to the investigated test compoundâ€™s potency. [Figure not available: see fulltext.]

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